Optimalization of deoxyribonucleic acid extraction using various types of magnetic particles

0504208 - ÚMCH 2020 RIV SK eng J - Journal Article
Konečná, J. - Romanovská, D. - Horák, Daniel - Trachtová, Š.
Optimalization of deoxyribonucleic acid extraction using various types of magnetic particles.
Chemical Papers. Roč. 73, č. 5 (2019), s. 1247-1255. ISSN 2585-7290. E-ISSN 1336-9075
Institutional support: RVO:61389013
Keywords : magnetic particles * solid-phase extraction * polymerase chain reaction
OECD category: Polymer science
Impact factor: 1.680, year: 2019
Method of publishing: Limited access
https://link.springer.com/article/10.1007%2Fs11696-018-00675-9

Isolation of deoxyribonucleic acid is an important step in the molecular diagnostics of microorganisms. A high quality of isolated deoxyribonucleic acid is necessary for deoxyribonucleic acid amplification by the polymerase chain reaction. The conventional deoxyribonucleic acid isolation using phenol–chloroform extraction and deoxyribonucleic acid precipitation in ethanol is time-consuming and requires the use of toxic phenol. Magnetic separation techniques using magnetic solid particles are one of modern methods to speed up the nucleic acids isolation. The aim of this work was to use two different types of magnetic particles for solid-phase deoxyribonucleic acid extraction. The amounts of deoxyribonucleic acid in separation mixtures were measured using ultraviolet spectrophotometry. The first experimental conditions were tested on chicken erythrocytes deoxyribonucleic acid. Phosphate buffer (pH 7, 7.6 and 8) was used for the adsorption of deoxyribonucleic acid on magnetic particles. Tested values of pH have no effects on deoxyribonucleic acid adsorption. It was shown that approximately almost one half of deoxyribonucleic acid was adsorbed to the particles. A number of different elution conditions (temperature, time and pH value of elution buffer) were investigated to determine their effect on elution efficiency. It was shown that only a small fraction of the bound DNA was released at 22 °C, while the release was more effective as the temperature was increased also the amount of elution deoxyribonucleic acid was more effective when time was increased. The higher amounts of deoxyribonucleic acid were eluted with TE buffer pH 9.0. Second, bacterial deoxyribonucleic acid was tested. This deoxyribonucleic acid eluted from the particles was in polymerase chain reaction ready quality.
Permanent Link: http://hdl.handle.net/11104/0295934